Objective To investigate the expression of runt-related transcription factor 1 translocated to 1 (RUNX1T1) and neuronal differentiation in hippocampal radial glial cells after deafferented injury of hippocampus in vitro.  Methods After physical transecting of fimbriafornix in 6 rats to establish a model of deafferented hippocampus, the extract of hippocampus was prepared. Hippocampal radial glial cells were cultured into 6 pieces of 24 holes cell culture plates in vitro and divided into deafferented injury, normal and blank groups, 48 holes in each group. The expression of RUNX1T1 and proportion of microtubule associated protein 2 (MAP-2) positive neurons in each group were detected by immunofluorescence, Real-time PCR and Western blotting.   Results Normally hippocampal radial glial cells had thin and long processes and expressed RUNX1T1. The fluorescence intensity of RUNX1T1 in deafferented injury group was about 1.8 times higher than that in normal and blank group, and the cell process was also longer. The mRNA and protein expression of RUNX1T1 in the deafferented injury group were increased by 2.9 and 2.4 times respectively, and 39.33% cells expressed MAP-2. Compared with normal and blank group, the proportion of MAP-2 positive neurons increased more.   Conclusion The expression of RUNX1T1 is up-regulated in hippocampal radial glial cells after deafferented injury, and they could differentiate into more neurons in vitro.